https://scholars.lib.ntu.edu.tw/handle/123456789/162631
標題: | 研究第二型登革病毒血清型專一性抗體的B細胞抗原決定位及評估其為偵檢試劑之可行性 | 作者: | 吳漢忠 | 關鍵字: | 第二型登革病毒;噬菌體顯現法;抗原決定位;登革偵檢試劑;monoclonal antibody;B-cell epitope;synthetic peptide;serologic diagnosis | 公開日期: | 2003 | 出版社: | 臺北市:國立臺灣大學醫學院口腔生物科學研究所 | 摘要: | 本計畫主要是要研究登革病毒的B 細 胞抗原決定位。經由ELISA, immunofluorescence 和immunoblotting 方 法,我們找出具專一性的第二型登革病毒 單株抗體(Ab4)。此抗體之抗原決定位的確 認,有助於病毒感染機制的暸解,疫苗及 偵檢試劑的研發。在此研究中,我們以噬 菌體顯現法來研究第二型登革病毒的抗原 決定位。我們也已經篩選出多株的噬菌體 可以專一性的與第二型登革病毒抗體結 合。這些專一性噬菌體所攜帶的外來 peptide,都含有His-Arg/Lys-Leu/Ile motif。 以synthetic peptide 代替噬菌體所攜帶的外 來peptide,可與第二型登革病毒抗體明顯 反應。第二型登革病毒感染之老鼠及兔子 血清,可以用此epitope-based peptide 來確 認。另外,此epitope-based peptide 可有效 的分辨登革病毒及日本腦炎病毒感染之老 鼠血清,這些抗原決定位的確認與研究, 將有助於epitope-based peptide antigen 之研 發,及了解出血性登革之病理機制。 Serotype-specific monoclonal antibody (MAb), D2 16-1 (Ab4), against dengue virus type 2 (DEN-2) was generated. This MAb recognized non-structural protein 1 (NS1) of DEN-2; its specificity was determined by ELISA, immunofluorescence and immunoblotting analysis. Serotype-specific B-cell epitope of Ab4 was further identified from a random phage-displayed peptide library. Selected phage clones specifically reacted with Ab4 and did not react with other MAbs. Immunopositive phage clones displayed a consensus motif, His-Arg/Lys-Leu/Ile. Synthetic peptide corresponding to phage-borne peptide specifically bound to antibody. His and Arg in this epitope were crucial for peptide binding to Ab4: binding activity dramatically decreased when His or Arg was changed to Leu. Epitope-based synthetic peptide identified serum samples from DEN-2-immunized mice and rabbits by ELISA assay. The peptide clearly differentiated between serum samples from DEN-2- and Japanese encephalitis virus (JEV)-immunized mice. This monoclonal antibody and its epitope-based peptide antigen will be useful for serologic diagnosis of DEN-2 infection. Furthermore, DEN-2 epitope identification makes it feasible to dissect the antibody response and to address the role of antibodies in the pathogenesis of primary and secondary DEN-2 infections. |
URI: | http://ntur.lib.ntu.edu.tw//handle/246246/22667 | 其他識別: | 912314B002257 | Rights: | 國立臺灣大學醫學院口腔生物科學研究所 |
顯示於: | 口腔生物科學研究所 |
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912314B002257.pdf | 97.51 kB | Adobe PDF | 檢視/開啟 |
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